CN111187364A - Ultrahigh pressure extraction method of lentinan - Google Patents
Ultrahigh pressure extraction method of lentinan Download PDFInfo
- Publication number
- CN111187364A CN111187364A CN201811356062.0A CN201811356062A CN111187364A CN 111187364 A CN111187364 A CN 111187364A CN 201811356062 A CN201811356062 A CN 201811356062A CN 111187364 A CN111187364 A CN 111187364A
- Authority
- CN
- China
- Prior art keywords
- lentinan
- ethanol
- ultrahigh pressure
- adsorption
- concentrating
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Sustainable Development (AREA)
- Biochemistry (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention relates to a method for extracting lentinan. The invention applies ultrahigh pressure technology, which is a processing method that the pressure is more than 100MPa to the raw material under the conditions of normal temperature and near normal temperature, the ultrahigh pressure can lead the solvent to enter the interior of plant cells, the solvent can overflow with effective components after pressure relief, high extraction efficiency of the effective components of the plant is realized, the lentinan is adsorbed and purified by utilizing macroporous resin, the adsorption selectivity to the lentinan is good, the adsorption speed and the resolution are fast, the adsorption capacity is larger, and the defects of relatively lower extraction rate, low extraction purity and the like of the conventional technology are overcome.
Description
The technical field is as follows:
the invention belongs to the field of natural organic chemistry, relates to a lentinan purification method, and particularly relates to a method for extracting lentinan from lentinus edodes by an ultrahigh pressure technology.
Background art:
lentinus edodes (academic name: Lentinuseedodes), also called Lentinus edodes, Pleurotus ostreatus, Mycopharyngodon comatus, Pleurotus ostreatus, Flammulina velutipes and Lentinus edodes, is an edible fungus. The edible part is the fruiting body of the mushroom, and the dried mushroom is obtained by dehydrating the fresh mushroom, so that the mushroom is convenient to transport and store. The edible part of the dried mushroom accounts for 72 percent, and each 100g of the edible part contains 13g of water, 1.8g of fat, 54g of carbohydrate, 7.8g of crude fiber, 4.9g of ash, 124mg of calcium, 415mg of phosphorus, 25.3mg of iron, 10.07mg of vitamin B, 21.13mg of vitamin B and 18.9mg of nicotinic acid. The Lentinus Edodes contains abundant lentinan, and can improve the activity of helper T cell to enhance humoral immunity function of human body. A large number of practices prove that the shiitake has a wide cancer prevention and treatment range and is used for clinical treatment. The Lentinus Edodes has effects of promoting metabolism, improving organism adaptability, treating diabetes, pulmonary tuberculosis, infectious hepatitis, neuritis, and can be used for treating dyspepsia, constipation, and reducing weight. Modern researches prove that lentinan can regulate the activity of T cells with immune functions in human bodies and can reduce the tumor inducing capacity of methylcholanthrene. The mushroom has strong inhibition effect on cancer cells, the inhibition rate on the mouse sarcoma 180 is 97.5%, and the inhibition rate on the ehrlich cancer is 80%. The shiitake mushroom also contains double-stranded ribonucleic acid, can induce to generate interferon, and has antiviral ability.
Lentinan is one of the most effective components of lentinus edodes, and is glucan which is composed of three strands of monosaccharide chains and has a spiral stereo configuration (tertiary structure), the stereo configuration of the glucan is similar to deoxyribonucleic acid (DNA) and ribonucleic acid (RNA), the lentinan is a macromolecular compound, the molecular weight of the glucan is thousands to hundreds of thousands, the glucan is insoluble in high-concentration alcohol, slightly soluble in low-concentration alcohol and cold water, and the glucan can be completely dissolved in hot water. Lentinan is present on the inner wall of the cell wall of lentinus edodes. The conventional technology for lentinan at present has the defects of relatively low extraction rate, low extraction purity and the like.
The invention content is as follows:
the invention aims to overcome the defects of relatively low extraction rate, low extraction purity and the like of the conventional technology and provides a method for extracting lentinan from lentinus edodes by using an ultrahigh pressure technology.
In order to achieve the purpose, the invention adopts the technical scheme that:
a method for extracting lentinan from lentinus edodes by using an ultrahigh pressure technology comprises the following steps:
(1) crushing dried shiitake mushrooms, sieving the crushed shiitake mushrooms with a 60-80-mesh sieve, putting the crushed shiitake mushrooms into a pressure-resistant bag, adding 10-12 times of ethanol with the volume concentration of 70% -90%, treating the mixture under the conditions of ultrahigh pressure of 200MPA and 300MPA for 10-15min at 18-20 ℃, recovering the ethanol, and drying the mixture in vacuum at 60 ℃ to obtain a crude product;
(2) dissolving the obtained lentinan crude product powder with a proper amount of 75% ethanol, adjusting the pH value to 3.0-7.5, heating to 40-60 ℃, adding 1.0-2.0% of pectinase (100 ten thousand u/ml), uniformly mixing, and keeping enzymolysis for 0.5-3 h. Cooling to room temperature, standing, filtering, discarding residue, and concentrating the filtrate under reduced pressure to obtain lentinan crude extract;
(3) adsorbing the obtained lentinan crude extract by using macroporous adsorption resin;
(4) eluting macroporous adsorbent resin with ethanol of different concentrations, tracking and detecting with thin layer chromatography, and collecting eluate of lentinan at each stage;
(5) concentrating the eluate of lentinan at each stage, and recovering ethanol to obtain lentinan semi-finished product with content of above 40%;
(6) loading the obtained semi-finished product of lentinan onto silica gel chromatographic column, eluting with n-butanol and chloroform at different volume ratios, tracking and detecting by thin layer chromatography, and collecting eluate of lentinan at each stage;
(7) dissolving the product with 6-8 times volume of 60-80% ethanol, concentrating to one tenth of the original volume, standing overnight to obtain crystal; separating crystal, dissolving the residue with 50% -60% ethanol, concentrating again, recrystallizing, and repeating the above steps twice to obtain refined lentinan with content of above 90%.
The macroporous absorption resin used in the step (3) is X-5, NKA-9, S-8, D3520, D4006, H103, D4020, AB-
One of 8H and LX-100 type resins, the adsorption is static adsorption or dynamic adsorption.
And (4) eluting by using 70-90% ethanol.
The invention applies the ultrahigh pressure technology which is a processing method for applying pressure greater than 100 hydrostatic pressure to raw materials at normal temperature and near normal temperature, the ultrahigh pressure can enable a solvent to enter plant cells, the solvent can overflow with effective components after pressure relief, high extraction efficiency of the effective components of the plant is realized, the lentinan is adsorbed and purified by using macroporous resin, the adsorption selectivity to the lentinan is good, the adsorption speed and the resolution are fast, and the adsorption capacity is larger;
the invention selects the macroporous adsorption resin with stable physicochemical property, larger surface area, higher exchange speed, high mechanical strength, strong pollution resistance and good thermal stability, is not compatible with acid, alkali and organic medium, has better selectivity to organic matters, is not influenced by the existence of inorganic salts and strong ion low molecular compounds, can selectively adsorb the lentinan from the solution by physical adsorption, and has fast adsorption, fast analysis and larger adsorption capacity.
The specific implementation mode is as follows:
example 1
A method for extracting lentinan from lentinus edodes by using an ultrahigh pressure technology comprises the following steps:
(1) pulverizing dried Lentinus Edodes, sieving with 60 mesh sieve, placing into pressure-resistant bag, adding 10 times volume of 70% ethanol, treating under ultrahigh pressure of 200MPA at 18 deg.C for 10min, recovering ethanol, and vacuum drying at 60 deg.C to obtain crude product;
(2) dissolving the obtained lentinan crude powder with appropriate amount of 75% ethanol, adjusting pH to 5.5, heating to 60 deg.C, adding pectase (100 ten thousand u/ml) 2.0%, mixing, and maintaining enzymolysis for 0.5 h. Cooling to room temperature, standing, filtering, discarding residue, and concentrating the filtrate under reduced pressure to obtain lentinan crude extract;
(3) adsorbing the obtained lentinan crude extract with LX-100 type macroporous adsorbent resin;
(4) eluting macroporous adsorbent resin with 70%, 80% and 90% ethanol, detecting by thin layer chromatography, and collecting eluate of lentinan at each stage;
(5) concentrating the eluate of lentinan at each stage, and recovering ethanol to obtain semi-finished product of lentinan with content of 46.3%;
(6) subjecting the obtained semi-finished product of lentinan to silica gel chromatography column, eluting with n-butanol and chloroform at ratio of 1: 3, 1: 4, 1: 5, 1: 6, and 1: 7, tracking and detecting by thin layer chromatography, and collecting eluate of lentinan at each stage;
(7) dissolving the product with 60% ethanol 6 times the volume of the product, concentrating to one tenth of the original volume, and standing overnight to obtain crystals; separating crystal, dissolving the residue with 50% ethanol, concentrating again, recrystallizing, and repeating the above steps twice to obtain refined lentinan with content of more than 92.5%.
Example 2
A method for extracting lentinan from lentinus edodes by using an ultrahigh pressure technology comprises the following steps:
(1) pulverizing dried Lentinus Edodes, sieving with 80 mesh sieve, placing into pressure-resistant bag, adding 12 times volume of 90% ethanol, processing under ultrahigh pressure of 300MPA at 20 deg.C for 15min, recovering ethanol, and vacuum drying at 60 deg.C to obtain crude product;
(2) dissolving the obtained lentinan crude powder with appropriate amount of 75% ethanol, adjusting pH to 7.5, heating to 60 deg.C, adding pectase (100 ten thousand u/ml) 2.0%, mixing, and maintaining enzymolysis for 1.5 h. Cooling to room temperature, standing, filtering, discarding residue, and concentrating the filtrate under reduced pressure to obtain lentinan crude extract;
(3) adsorbing the obtained lentinan crude extract with AB-8H macroporous adsorbent resin;
(4) eluting macroporous adsorbent resin with 70%, 80%, 90% ethanol, tracking and detecting with thin layer chromatography, and collecting eluate of lentinan at each stage;
(5) concentrating the eluate of lentinan at each stage, and recovering ethanol to obtain lentinan semi-finished product with content of above 41.4%;
(6) subjecting the obtained semi-finished product of lentinan to silica gel chromatography column, eluting with n-butanol and chloroform at ratio of 1: 3, 1: 4, 1: 5, 1: 6, and 1: 7, tracking and detecting by thin layer chromatography, and collecting eluate of lentinan at each stage;
(7) dissolving the product with 80% ethanol 8 times the volume of the product, concentrating to one tenth of the original volume, and standing overnight to obtain crystals; separating crystal, dissolving the residue with 60% ethanol, concentrating again, recrystallizing, and repeating twice to obtain refined lentinan with content of 91.7%.
Embodiment 3
A method for extracting lentinan from lentinus edodes by using an ultrahigh pressure technology comprises the following steps:
(1) pulverizing dried Lentinus Edodes, sieving with 80 mesh sieve, placing into pressure-resistant bag, adding 80% ethanol 12 times the volume of Lentinus Edodes, processing under ultrahigh pressure of 200MPA at 18 deg.C for 12min, recovering ethanol, and vacuum drying at 60 deg.C to obtain crude product;
(2) dissolving the obtained lentinan crude powder with appropriate amount of 75% ethanol, adjusting pH to 4.5, heating to 50 deg.C, adding 1.5% pectase (100 ten thousand u/ml), mixing, and maintaining enzymolysis for 2 hr. Cooling to room temperature, standing, filtering, discarding residue, and concentrating the filtrate under reduced pressure to obtain lentinan crude extract;
(3) adsorbing the obtained lentinan crude extract by using H103 macroporous adsorption resin;
(4) eluting macroporous adsorbent resin with 70%, 80%, 90% ethanol, and thin layer chromatography
Tracking and detecting, and collecting the eluents of each stage of lentinan;
(5) concentrating the eluate of lentinan at each stage, and recovering ethanol to obtain lentinan semi-finished product with content of 42.6%;
(6) subjecting the obtained semi-finished product of lentinan to silica gel chromatography column, eluting with n-butanol and chloroform at ratio of 1: 3, 1: 4, 1: 5, 1: 6, and 1: 7, tracking and detecting by thin layer chromatography, and collecting eluate of lentinan at each stage;
(7) dissolving the product with 70% ethanol 8 times the volume of the product, concentrating to one tenth of the original volume, and standing overnight to obtain crystals; separating crystal, dissolving the residue with 60% ethanol, concentrating again, recrystallizing, and repeating twice to obtain refined lentinan with content of 93.6%.
Claims (3)
1. A method for extracting lentinan from lentinus edodes by using an ultrahigh pressure technology comprises the following steps:
(1) crushing dried shiitake mushrooms, sieving the crushed shiitake mushrooms with a 60-80-mesh sieve, putting the crushed shiitake mushrooms into a pressure-resistant bag, adding 10-12 times of ethanol with the volume concentration of 70% -90%, treating the mixture under the conditions of ultrahigh pressure of 200MPA and 300MPA for 10-15min at 18-20 ℃, recovering the ethanol, and drying the mixture in vacuum at 60 ℃ to obtain a crude product;
(2) dissolving the obtained lentinan crude product powder with a proper amount of 75% ethanol, adjusting the pH value to 3.0-7.5, heating to 40-60 ℃, adding 1.0-2.0% of pectinase (100 ten thousand u/ml), uniformly mixing, and keeping enzymolysis for 0.5-3 h;
cooling to room temperature, standing, filtering, discarding residue, and concentrating the filtrate under reduced pressure to obtain lentinan crude extract;
(3) adsorbing the obtained lentinan crude extract by using macroporous adsorption resin;
(4) eluting macroporous adsorbent resin with ethanol of different concentrations, tracking and detecting with thin layer chromatography, and collecting lentinus edodes
Eluent in each stage of sugar;
(5) concentrating the eluate of lentinan at each stage, and recovering ethanol to obtain lentinan semi-finished product with content of above 40%;
(6) loading the obtained semi-finished product of lentinan onto silica gel chromatographic column, eluting with n-butanol and chloroform at different volume ratios, tracking and detecting by thin layer chromatography, and collecting eluate of lentinan at each stage;
(7) dissolving the product with 6-8 times volume of 60-80% ethanol, concentrating to one tenth of the original volume, standing overnight to obtain crystal; separating crystal, dissolving the remainder with 50% -60% ethanol, concentrating again, recrystallizing, repeating twice to obtain refined lentinan with content of above 90%;
the macroporous adsorption resin used in the step (3) is one of resins of X-5, NKA-9, S-8, D3520, D4006, H103, D4020, AB-8H and LX-100, and the adsorption is static adsorption or dynamic adsorption;
and (4) eluting by using 70-90% ethanol.
2. The method for extracting lentinan by using the ultrahigh pressure technology as claimed in claim 1, which is characterized in that: in the step (1), the mushroom powder is sieved by a sieve of 60-80 meshes and then is subjected to ultrahigh pressure extraction.
3. The method for extracting lentinan by using the ultrahigh pressure technology as claimed in claim l, which is characterized in that: in the step (2), after the ultrahigh pressure extraction is finished, the enzymolysis technology is utilized for re-extraction.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811356062.0A CN111187364A (en) | 2018-11-15 | 2018-11-15 | Ultrahigh pressure extraction method of lentinan |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811356062.0A CN111187364A (en) | 2018-11-15 | 2018-11-15 | Ultrahigh pressure extraction method of lentinan |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111187364A true CN111187364A (en) | 2020-05-22 |
Family
ID=70703219
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811356062.0A Pending CN111187364A (en) | 2018-11-15 | 2018-11-15 | Ultrahigh pressure extraction method of lentinan |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111187364A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113940420A (en) * | 2020-07-18 | 2022-01-18 | 哈尔滨腾凝科技有限公司 | Method for promoting release of mushroom flavor substances by using ultrahigh pressure wall breaking technology |
| CN114487173A (en) * | 2022-01-13 | 2022-05-13 | 杭州傲敏生物科技有限公司 | High-efficiency liquid-phase fingerprint spectrum identification method for lentinus edodes powder polysaccharide |
-
2018
- 2018-11-15 CN CN201811356062.0A patent/CN111187364A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113940420A (en) * | 2020-07-18 | 2022-01-18 | 哈尔滨腾凝科技有限公司 | Method for promoting release of mushroom flavor substances by using ultrahigh pressure wall breaking technology |
| CN114487173A (en) * | 2022-01-13 | 2022-05-13 | 杭州傲敏生物科技有限公司 | High-efficiency liquid-phase fingerprint spectrum identification method for lentinus edodes powder polysaccharide |
| CN114487173B (en) * | 2022-01-13 | 2023-11-28 | 杭州傲敏生物科技有限公司 | High-efficiency liquid-phase fingerprint identification method for lentinan |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101157712B (en) | Method for separating and purifying cordycepin | |
| CN109694416B (en) | Preparation method of high-purity, high-solubility and high-activity edible fungus polysaccharide | |
| CN102838686B (en) | Method for preparing high-purity agaricus blazei murrill polysaccharide | |
| CN105037573B (en) | A kind of preparation and its application with antiviral and immunocompetent Linseed pigment | |
| CN111187364A (en) | Ultrahigh pressure extraction method of lentinan | |
| CN102078339B (en) | Method for enriching and purifying common phellinus fungus general flavone in common phellinus fungus | |
| CN103497258A (en) | Method for extracting and purifying active polysaccharides of wheat bran | |
| CN105053952B (en) | A kind of processing technology of the dried orange peel extracts of no bitter taste | |
| CN101074188A (en) | Method for enriching and purifying veralkcohol from peanut root by macporous adsorptive resin | |
| CN103494858A (en) | Method for enriching total flavonoids in taraxacum mongolicum with macroporous resins | |
| CN102093328B (en) | Method for enriching and purifying procyanidin in pine bark | |
| CN103408610A (en) | Method for extracting arbutin from pear leaves | |
| CN105566409A (en) | Method for extracting and separating glucoraphanin from broccoli seeds | |
| CN102432656B (en) | Method for extracting and purifying cordycepin from Cordyceps militaris sporophore | |
| CN103059086A (en) | Extraction and purification method of cordycepin from cordyceps militaris solid mediums | |
| CN102936271B (en) | Method for extracting cordycepin | |
| CN111116766A (en) | Method for extracting lycium barbarum polysaccharide by using ultrahigh pressure technology | |
| CN113440547B (en) | Method for separating and purifying Japanese thistle herb total glycosides by adopting macroporous resin series dynamic axial compression column | |
| CN101805269A (en) | Method for separating and extracting natural theanine | |
| CN111217780A (en) | Method for extracting atractylone by using ultrahigh pressure technology | |
| CN101921346B (en) | Radial flow chromatography for polysaccharides of mushroom hyphae | |
| CN103387501A (en) | Method for preparing high-purity L-synephrine | |
| CN104961667A (en) | Purifying method of sulforaphane | |
| CN111419892A (en) | Extraction method of abrus herb total flavone | |
| CN104672090A (en) | Method for separating and purifying ramie leaf chlorogenic acid |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20200522 |